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  • 1
    ISSN: 0952-3499
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A secreted form of phospholipase A2 (PLA2) is thought to play an important role in inflammatory diseases. To characterize this enzyme the cDNA encoding a low molecular weight PLA2 was cloned from a human placental cDNA library. The cDNA encoding the human PLA2 was subcloned into an expression vector and subsequently transfected into Chinese hamster ovary (CHO) cells. A stable CHO cell clone, secreting ca I mg/L of recombinant PLA2 into the medium, was scaled up in culture to 180L. The recombinant enzyme was purified from the cell supernatant to apparent homogeneity by a novel procedure combining adsorption to poly(vinylidene difluoride) membranes, ion exchange chromatography and size exclusion chromatography. The final recovery of PLA2 activity was 58%. A direct comparison between the purified recombinant human PLA2 and PLA2 purified from human synovial fluid, including molecular weight, antigenicity, ionic dependence, substrate specificity and sensitivity to know PLA2 inhibitors, indicated that the two enzymes exhibit identical biochemical properties. These results show that the recombinant PLA2 can be efficiently expressed and purified in sufficient quantities to characterize the enzyme active site, to aid in the rational development of PLA2 inhibitors as potential anti-inflammatory drugs, and to investigate further the role of PLA2 in inflammatory disease.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Chichester : Wiley-Blackwell
    Journal of Chemical Technology AND Biotechnology 59 (1994), S. 115-115 
    ISSN: 0268-2575
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Biochemical and Microbiological Technology and Engineering 1 (1959), S. 77-98 
    ISSN: 0368-1467
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Fractions of markedly different protein contents may be obtained by air-classifying flour (in the sub-sieve range) at cut sizes of about 17 and 35 μ respectively. Of these fractions, the fine, 1 (of high protein content), contains fragments of interstitial endosperm protein and small starch granules; the intermediate, 2 (low protein), mainly large starch granules; the coarse, 3, fragments of endosperm cells. The yields and protein contents of the fractions vary with different wheats; the yields of 1 and 2 may be greatly increased through suitably grinding the flour after milling.In general, 1 may be used for raising the protein content of flours deficient in protein to a level suitable for bread-making. The amount of 1 required depends on its protein contents and on whether it came from a hard or soft wheat, the hard wheat fraction having better bread-making qualities. The resulting variation in the proportion of 1 required in the blend causes variation in the proportion of other particulate materials introduced with 1, in which they have become concentrated during the air classification. These include diastatically active agents and finely divided discolouring matter; the level of the former reached in the blend greatly affects its bread-making value. Pre-grinding of the flour may be helpful in this respect.When chlorinated, 2 is potentially useful for making light-structured cakes, but hard English wheats have mainly given less satisfactory results than soft. Under certain conditions, 3, with or without 2, may be better for biscuit-making than the parent flour.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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